Single locus microsatellites isolated using 5' anchored PCR

Fisher, P. J.; Gardner, Richard C.; Richardson, Thomas E.

doi:10.1093/nar/24.21.4369

Cited by 144 publications

(106 citation statements)

References 9 publications

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“…Molecular markers based on SSR (Simple Sequence Repeats) are powerful techniques because they are codominant, multi-allelic, easily scored and highly polymorphic, which can be used to identify and determine plant genomes or to evaluate the phylogenetic relationship among genotypes of barley [4] [5] [6]. However, a major drawback of microsatellite markers is the time and cost required to characterize them [7].…”

Section: Introductionmentioning

confidence: 99%

Efficiency of Triple-SCoT Primer in Characterization of Genetic Diversity and Genotype-Specific Markers against SSR Fingerprint in Some Egyptian Barley Genotypes

Aboulila¹,

Mansour²

2017

AJMB

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Ten Egyptian barley genotypes (2 commercial varieties and 8 breeding lines) were cultivated under normal condition at the Experimental Farm of Sakha Agricultural Research station and exposed to salinity stress condition at the Experimental Farm of El-hosainia plain Agricultural Research station, Elsharkia Governorate, Egypt, in an attempt to identify the relative salinity tolerant genotypes. A susceptibility index (SI) was used to estimate the relative stress loss because it accounted for variation in yield potential and stress intensity. Giza 123, Line-1, Line-5, Line-6 and Line-8 genotypes were considered as saline tolerant genotypes on the basis of their highly tolerance indices values. Barley genotypes were characterized by seven SSR markers and three SCoT primers in different combinations to discern the extent of genetic variation and develop a fingerprinting key. Normal SCoT reactions amplify single segments of DNA which are 15-to 19-mer long. A new strategy was used to increase SCoT potential in genetic diversity studies by using two and three different primer combinations per reaction. Amplification products scored a polymorphism percentage of 94.44% for Triple-SCoT and 90.91% for SSR, while the average no. of polymorphic fragments/primer was 17 and 7.14 in the two marker systems, respectively. On the other side, Triple-SCoT exhibited the highest average number of positive and negative genotype-specific markers. The cluster analysis of the studied genotypes using these different marker systems revealed four dendrograms varied in their topology. The dendrogram based on Triple-SCoT data exhibited the closest relationships to those illustrated by SSR dendrogram.

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Section: Introductionmentioning

confidence: 99%

Efficiency of Triple-SCoT Primer in Characterization of Genetic Diversity and Genotype-Specific Markers against SSR Fingerprint in Some Egyptian Barley Genotypes

Aboulila¹,

Mansour²

2017

AJMB

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“…A PCR-based library that was enriched for different microsatellite motifs was constructed by the 5ꞌ-anchored PCR technique (Fisher et al, 1996). Briefly, the sequences that contained microsatellite motifs were amplified by a single primer anchored at the 5ꞌ-end of microsatellite by seven degenerate bases (PCT6: 5ꞌ-KKVRVRV (CT) 6 , K = G/T, V = G/C/A, R = G/A).…”

Section: Methodsmentioning

confidence: 99%

Rapid isolation and characterization of polymorphic microsatellite loci in the mud crab, Scylla paramamosain (Portunidae)

Yao

Sun²,

Wang³

et al. 2012

Genet. Mol. Res.

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“…Microsatellites were isolated and characterized using 5 anchored PCR primers 21 and partial enriched library as modified from Hamilton et al 22 . Microsatellite primers were designed using PRIMER 3 23 .…”

Section: Dna Isolation and Microsatellite Analysismentioning

confidence: 99%

Microsatellite marker variation in populations of the melon fly parasitoid, Psyttalia fletcheri

Homchan¹,

Haymer²,

Kitthawee³

2014

ScienceAsia

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The parasitoid Psyttalia fletcheri (Silvestri) is an important natural enemy of the melon fly, Bactrocera cucurbitae (Coquillett). Melon fly infestations are responsible for extensive losses of cucurbit production worldwide, and P. fletcheri has been used for some time in biological control programmes attempting to deal with this pest. However, there is a general lack of knowledge of the genetic structure of populations of P. fletcheri, and the development of this information is key to the effective use of this parasitoid. In this study, we isolated several novel microsatellite loci to investigate the genetic structure of P. fletcheri populations from six locations in Thailand. All the loci analysed here were polymorphic, and the mean number of alleles per locus ranged from 4.2-8.6. Heterozygote deficiencies were noticed in most populations. Overall FST estimates showed moderate genetic differentiation among P. fletcheri populations with a jackknife mean of 0.084. However, pairwise FST calculations revealed that 11 out of 15 population comparisons showed genetic differentiation. The greatest level of differentiation was also found for the population that had the lowest value for genetic diversity. In contrast, populations with high levels of genetic variation did not show significant genetic differentiation, nor did they show significant isolation by distance. An unrooted dendrogram constructed from Nei's genetic distance values also confirmed that one population from the south of Thailand can be separated from the others.

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Single locus microsatellites isolated using 5' anchored PCR

Cited by 144 publications

References 9 publications

Efficiency of Triple-SCoT Primer in Characterization of Genetic Diversity and Genotype-Specific Markers against SSR Fingerprint in Some Egyptian Barley Genotypes

Efficiency of Triple-SCoT Primer in Characterization of Genetic Diversity and Genotype-Specific Markers against SSR Fingerprint in Some Egyptian Barley Genotypes

Rapid isolation and characterization of polymorphic microsatellite loci in the mud crab, Scylla paramamosain (Portunidae)

Microsatellite marker variation in populations of the melon fly parasitoid, Psyttalia fletcheri

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